Effect of temperature on seed germination of ten species and varieties ofLimnanthes

Ten accessions of seed of the genus Limnanthes, representing seven species and their varieties, were tested for germination at temperatures of 40°, 50°, 60°, 70° F, and at room temperature varying from 72°–78° F. Four of the ten accessions germinated best at 40°, three at 50°, and three at 60° F. Germination was relatively poor at 70° F and almost zero at room temperature. The total germination averaged over all accessions was about the same for both 40° F and 50° F, slightly less for 60° F. Seeds at 60° F reached a peak in germination rate in four days; those at 50° F required six days; those at 40° F required 11 days. Seeds, started at room temperature and moved to 50° F 15 days later, attained only about half the total germination of a similar group of seeds originlly started at 50° F. Some of the species tested germinated almost equally well at 40°, 50°, or 60° F; others germinated best only at, or near, one of these temperatures. Substantially all of the seeds that germinated over a period of 50 days did so in less than 30 days; most of these germinated in the first 14 days.     

Germination traits and seed-bank dynamics of a biennial plant,Oenothera glazioviana Micheli

A study was conducted on the germination traits and seed-bank dynamics ofOenothera glazioviana (=O. erythrosepala), which sets seed in August in sand-dune systems in Japan. More than 90% of freshly matured seeds germinated over a wide range of temperature in light, but less than 10% did so in continuous darkness. Stratification (chilling under moist conditions) was ineffective in diminishing the light-requirement for germination. When fresh seeds were imbibed for 24 h including a 12-h light period, followed by 7-day air-drying, 94% of them became germinable in the dark at 25°C, but remained dormant at less than 15°C. of seeds collected in March from capsules of dead plants, 58% germinated in the dark at 25°C. After four cycles of alternatc 1-day wetting followed by 2-day drying or 1.5-day wetting followed by 1.5-day drying under a 12-h photoperiod, the fraction of viable seeds declined from 76% to 40% and 22%, respectively, due to germination during the wet periods. Seed-bag experiments were conducted in the field, using seeds given and not given a light-stimulus. Forty percent of the light-stimulated seeds germinated in the soil, whereas the seeds without a light-stimulus remained dormant throughout the experiment. When seeds were placed on the soil surface or at a depth of 0.5-1 cm, the proportion of germinable seeds declined during late spring and autumn, but not during winter and early spring. The seed-bank size of a natural population just prior to current seed dispersal was 2–3% of the seed production in the previous year, suggesting a high turnover rate of the seed-bank.     

Effect of Gibberellic Acid on Dark and Light Germination at Different Temperatures of Calluna, Ledum andRhododendron Seeds

The aim of the experiments was to study the effects of gibberellic acid (GAs) on the germination of Calluna vulgaris L., Ledum palustre L. and Rhododendron lapponicum (L.) Wahlenb. seeds under different environmental conditions. Under continuous light from white fluorescense tubes (3000 lux), untreated seeds of Calluna were partly dormant at all temperatures studied (9, 15, 21, 27, 27/9, 8/16 hours). Percentage of dormant seeds increased, however, with decreasing temperature, and it varied also from seed lot to seed lot. Untreated seeds of Ledum were dormant in light at 9° and 15°C but not at higher temperatures. Untreated seeds of Rhododendron were completely dormant in light at temperatures from 13° to 24°C. Seeds of all species were completely dormant in darkness both at 15° and 27°C. GA₃ stimulated greatly the germination of all species under all studied environmental conditions. The used concentrations (0.2–3.2 mM) gave nearly 100% germination in most cases. At 9°C the dormancy in some seed lots of Calluna and Ledum was only partly broken by the used concentrations of GA₃.     

Factors affecting the dormancy and germination of bleeding heart [Lamprocapnos spectabilis (L.) Fukuhara] seeds

• Information on the optimal conditions to promote the germination of Lamprocapnos spectabilis (L.) Fukuhara seeds is limited; consequently, this study was conducted to establish the requirements to break seed dormancy and promote germination.
• The selected seeds had morphophysiological dormancy and had not begun embryo development. To study the dormancy breaking and embryo development processes, seeds were subjected to constant or changing temperature treatments during moist stratification.
• High temperature and humidity resulted in vigorous embryo growth, with the longest embryos occurring after 1 month of incubation at 20 °C. At 4 °C, the seeds required incubation period of at least 3 months to germinate. Embryo growth and germination were higher with changing high and low temperatures than under a constant temperature, and changing temperatures also considerably changed the endogenous hormone levels, embryo development and germination. Bioactive gibberellin (GA) content was higher in seeds incubated at 20 °C for 1 month, then at 4 °C for 2 months. The content of endogenous abscisic acid in seeds subjected to the same treatment decreased by 97.6% compared with that of the untreated seeds.
• Embryo growth and seed germination require changing high and low temperatures; however, exogenous GA₃ could substitute for high temperatures, as it also causes accelerated germination. In this study, the seeds of L. spectabilis were identified as an intermediate simple type, a sub‐level of morphophysiologically dormant seeds.

     

Population dynamics of the shrub Acacia suaveolens (Sm.) Willd.: Fire and the transition to seedlings

Fire, through soil heating effects, causes flushes of seed germination in Acacia suaveolens. Optimal temperatures for germination are between 60 and 80°C for any duration, or up to 100°C for durations less than 1 h. Exposure to temperatures less than 60°C leaves seeds dormant and viable, whilst seed death occurs in increasing proportions with increasing exposure to temperatures greater than 80°C. A field study of temperatures in the soil under simulated burns showed that the innate seed dormancy in A. suaveolens would only be broken for seeds up to a depth of 1 cm in ‘cool’ or 4 cm in ‘hot’ burns. In the hot burns some of the seeds in the top 1 cm of the soil were killed by excessive heating. These simulated burns most resemble cool and moderate/high intensity wildfires, respectively. Seeds can emerge from depths up to 8 cm and, for any seeds buried deeper than this, the probability of emergence is progressively reduced down to nil at 14 cm. Seeds buried between 5 and 10 cm will be heated sufficiently to break their dormancy only in a very high intensity wildfire. Seeds buried between 5 and 10 cm deep mostly occur in nests of an ant, Pheidole sp. Field observations of emergent seedlings confirm that post-fire emergence is concentrated over a small range of soil depths directly related to the intensity and duration of heating that occurs, whilst occasional seedlings may appear from greater or lesser depths largely dependent upon the spatial heterogeneity of soil heating in natural fires.     

Variation in seed dormancy and germination among populations of Silybum marianum (Asteraceae)

Milk thistle (Silybum marianum) is a medicinal plant; however, lack of consistency in past dormancy studies has hindered propagation of this species from seeds. We tested the germination responses of freshly harvested and after-ripened (stored for 2 and 7 months; 25°C at 50% relative humidity) seeds from three populations (P1, P2 and P3) in Iran at varying constant or alternating temperatures, with or without GA₃ and in light and continuous darkness. No germination occurred in freshly harvested seeds incubated at any condition without GA₃ application, indicating that all the seeds were dormant. Seeds from P1 and P2, which developed under relatively dry, warm conditions, germinated over a wider range of temperatures after 2 months of dry storage, indicating type 6 of non-deep physiological dormancy (PD). Seeds from P3, which developed under relatively wet, cool conditions, incubated at constant temperatures (especially on GA₃), exhibited an increase in maximum temperature for germination, indicating type 1 of non-deep PD. Light improved germination of after-ripened seeds, and GA₃ application substituted for the light requirement for germination. This is the first report that environmental conditions during seed development may be correlated with differences in the type of non-deep PD. We conclude that milk thistle seeds are positively photoblastic and photodormant and the germination responses of after-ripened seeds from different populations are different under darkness. Therefore, the impacts of genetic differences and maternal effects on the induction of dormancy during seed development should be considered in attempts to domesticate this medicinal plant.     

Response of Amaranthus retroflexus L. seeds to gibberellic acid, ethylene and abscisic acid depending on duration of stratification and burial

Freshly harvested, dormant seeds of Amaranthus retroflexus were unable to germinate at 25 and 35 °C. To release their dormancy at the above temperatures, the seeds were stratified at a constant temperature (4 °C) under laboratory conditions or at fluctuating temperatures in soil or by outdoor burial in soil. Fully dormant, or seeds stratified or buried (2006/2007 and 2007/2008) for various periods were treated with exogenous gibberellic acid (GA₃), ethephon and abscisic acid (ABA). Likewise, the effects of these regulators, applied during stratification, on seed germination were determined. The results indicate that A. retroflexus seed dormancy can be released either by stratification or by autumn–winter burial. The effect of GA₃ and ethylene, liberated from ethephon, applied after various periods of stratification or during stratification, depends on dormancy level. GA₃ did not affect or only slightly stimulated the germination of non-stratified, fully dormant seeds at 25 and 35 °C respectively. Ethylene increased germination at both temperatures. Seed response to GA₃ and ethylene at 25 °C was increased when dormancy was partially removed by stratification at constant or fluctuating temperatures or autumn–winter burial. The response to GA₃ and ethylene increased with increasing time of stratification. The presence of GA₃ and ethephon during stratification may stimulate germination at 35 °C. Thus, both GA₃ and ethylene can partially substitute the requirement for stratification or autumn–winter burial. Both hormones may also stimulate germination of secondary dormant seeds, exhumed in September. The response to ABA decreased in parallel with an increasing time of stratification and burial up to May 2007 or March 2008. Endogenous GA_n, ethylene and ABA may be involved in the control of dormancy state and germination of A. retroflexus. It is possible that releasing dormancy by stratification or partial burial is associated with changes in ABA/GA and ethylene balance and/or sensitivity to these hormones.     

Seed germination ecology of the threatened endemic Iberian <Emphasis Type="Italic">Delphinium fissum</Emphasis> subsp. <Emphasis Type="Italic">sordidum</Emphasis> (Ranunculaceae)

Seeds of Delphinium fissum subsp. sordidum are physiologically dormant at maturity, with underdeveloped embryos; thus they have morphophysiological dormancy (MPD). The aims of this study were to determine the requirements for embryo growth, dormancy break and germination, to characterise the type of seed dormancy and to evaluate the effects of light, seed age, pollination mechanism, and inter-annual and inter-population variability on germinative ability. After 3 months of incubation at 5°C (cold stratification) in darkness conditions, the mean embryo length increased from 5.6 to 2.07 mm, with 76% of seeds germinating. Conversely, embryos of seeds incubated during 3 months at 20/7 or 28/14°C hardly grew and no germination was recorded. Since cold stratification was the only requirement for the loss of MPD, and both dry storage in laboratory conditions and warm stratification prior to cold stratification shortened the cold stratification period required for germination, it could be concluded that D. fissum subsp. sordidum seeds have intermediate complex MPD. Cold stratification and incubation in darkness conditions promoted higher germination percentages than those in light. In addition, germinative ability increased with seed age up to 8 months (reaching 96% at 5°C in darkness), showed a pronounced inter-annual and inter-population variability, as well as a significant decrease in seeds coming from pollination by geitonogamy. High temperatures (25/10 or 28/14°C) induced seeds to secondary dormancy, so seedling emergence in the greenhouse was restricted to February–March. The requirements for dormancy break and germination reflect an adaptation to trigger germination in late winter. This study is the first one to document a gradual increase in germination percentage with seed age for plant species with intermediate complex MPD.     

Annual dormancy cycles in buried seeds of shrub species: germination ecology of Sideritis serrata (Labiatae)

The germination ecology of Sideritis serrata was investigated in order to improve ex‐situ propagation techniques and management of their habitat. Specifically, we analysed: (i) influence of temperature, light conditions and seed age on germination patterns; (ii) phenology of germination; (iii) germinative response of buried seeds to seasonal temperature changes; (iv) temperature requirements for induction and breaking of secondary dormancy; (v) ability to form persistent soil seed banks; and (vi) seed bank dynamics. Freshly matured seeds showed conditional physiological dormancy, germinating at low and cool temperatures but not at high ones (28/14 and 32/18 °C). Germination ability increased with time of dry storage, suggesting the existence of non‐deep physiological dormancy. Under unheated shade‐house conditions, germination was concentrated in the first autumn. S. serrata seeds buried and exposed to natural seasonal temperature variations in the shade‐house, exhibited an annual conditional dormancy/non‐dormancy cycle, coming out of conditional dormancy in summer and re‐entering it in winter. Non‐dormant seeds were clearly induced into dormancy when stratified at 5 or 15/4 °C for 8 weeks. Dormant seeds, stratified at 28/14 or 32/18 °C for 16 weeks, became non‐dormant if they were subsequently incubated over a temperature range from 15/4 to 32/18 °C. S. serrata is able to form small persistent soil seed banks. The maximum seed life span in the soil was 4 years, decreasing with burial depth. This is the second report of an annual conditional dormancy/non‐dormancy cycle in seeds of shrub species.     

Seed germination requirements of Buck’s beard [Aruncus dioicus (Walter) Fernald (Rosaceae)]

Aruncus dioicus (Walter) Fernald (Rosaceae) is a perennial herbaceous plant whose young shoots are traditionally collected in the wild and consumed as a food in NE Italy. The aim of this study was to determine the germination requirements of its seeds in order to start its cultivation, and to assess the germination of six accessions of the species. Viability of seeds ranged from 86 to 97% in the various accessions. Germination rate was almost null in seeds of two accessions, and ranged from 10.5 to 37.3 in the other ones. The seed coat was permeable to water. Treatments with GA₃, KNO₃ and mechanical scarification did not enhance the germination, while the cold stratification treatment at 2 °C for different periods improved the germination rate and the mean germination time as compared with the untreated seeds. With 45 days of cold stratification, the germination rate and mean germination time (respectively, 90.1% and 7.7 dd) of seeds were different from those of the untreated seeds. Cold stratified seeds germinated under artificial light and did not germinate in the dark. Seeds of A. dioicus displayed an intermediate physiological dormancy, removable by a cold stratification treatment, requiring both light and cold conditions.     

Seed Treatment Optimizes Benefits of Seed Bank Storage for Restoration‐Ready Seeds: The Feasibility of Prestorage Dormancy Alleviation for Mine‐Site Revegetation

Dormant seeds of 18 species from 9 families covering a diverse range of seed dormancy syndromes and life histories from the southwest Australian biodiversity hotspot were assessed for germinability following storage at 15–25°C for 36 months. A total of 10 species with physical dormancy (PY) and 8 with either physiological dormancy (PD) or morphophysiological dormancy (MPD) were assessed as part of the study. Prior to storage, germination from dormant seeds was 1–27%, rising to 41–100% following specific dormancy‐breaking treatments. When seed dormancy was removed prior to storage for 36 months seeds from all species were found to maintain a nondormant state and germinate to a similar level to that observed at the beginning of the experiment (44–100%). Likewise, seeds that did not receive a prestorage dormancy‐breaking treatment maintained a dormant state (0–50% germination) and subsequently responded well to a dormancy‐breaking treatment immediately prior to germination assessment (49–99%). There were minimal differences in response to dormancy‐breaking treatments before and after 36 months storage (average 4–6% difference) and in the germination responses observed between both storage environments assessed (15°C/15% eRH or 15–25°C air dried). Based on these findings, storing seeds in a nondormant state does not alter germinability and this approach provides significant benefits to current seed‐based restoration programs through reduction of double handling and improved seed use efficiency.     

Germination stimulation in wild oats (Avena fatua L.) by synthetic strigol analogs and gibberellic acid

At concentrations of 0.01–1 mM, five synthetic multiring analogs of strigol were effective germination stimulants of intact and dehulled wild oat (Avena fatua L.) seeds. The effect was concentration-dependent and equaled or exceeded that produced by equimolar gibberellic acid (GA₃). The most effective strigol analog treatments induced 55–80% germination within 7 days in intact wild oat seeds and resulted in 63–86% germination and normal seedling growth over 14 days. Intact wild oat controls germinated 14% after 14 days. The stimulation of wild oat germination by these synthetic strigol analogs demonstrates that these compounds, initially developed as germination stimulants for the seeds of the parasitic weed, witchweed (Striga asiatica L. Kuntz.), have bioregulatory activity in dormant seeds of monocots, as well as dicots. None of the compounds tested significantly affected the germination of nondormant cultivated oat seeds (Avena sativa L.). The commonly used dispersal agent, Tween 20 (0.1%), was found to inhibit germination of cultivated oats, alone and in the presence of 2% acetone.     

Seed storage and germination in Kumara plicatilis,a tree aloe endemic to mountain fynbos in the Boland,south-western Cape,South Africa

Seed storage under appropriate conditions is a relatively inexpensive means of safeguarding plant genetic material for ex situ conservation. Post-storage germination trials are used to determine the viability of stored seeds, and hence the efficacy of the particular storage treatment. Kumara plicatilis (= Aloe plicatilis) is a tree aloe endemic to mountain fynbos in the Boland, south-western Cape. The viability and germination behaviour of K. plicatilis seeds were assessed for seeds stored for four and nine months at − 80 °C, 4 °C, 25 °C and under ambient conditions in a laboratory. Seeds were germinated under controlled conditions and germination rates and percentages determined. Ungerminated seeds were tested for viability using tetrazolium salt. Seed viability was not significantly reduced during storage. Seeds stored at − 80 °C for four and nine months exhibited the fastest germination rate overall (both 5.9 ± 0.3 weeks, mean ± S.E.), and slowest was for seeds stored under ambient conditions for four and nine months (both 7.8 ± 0.4 weeks). All seed lots showed similar percentage germination after four months of storage (78.0–90.4%). The highest percentage germination overall was for seeds stored at − 80 °C for four months (90.4%) and the lowest was for seeds kept at 4 °C and − 80 °C for nine months (39.2 and 39.6%, respectively). Respective percentage viability for ungerminated seeds in these two treatments was 82% and 87%, respectively, indicating the induction of secondary dormancy. Induced dormancy triggered by protracted cold temperatures may be an adaptation that enables seeds to survive prolonged extreme conditions that are unfavourable for germination. Further research on the long-term storage of aloe seeds would be beneficial for developing long-term seed storage and germination testing protocols for ex situ conservation.     

INTERACTION OF TEMPERATURE AND GIBBERELLIN ON POTATO SEED GERMINATION

Recently harvested true seed of potato frequently requires several months for complete germination. Gibberellic acid (GA₃) has a pronounced effect on seed germination, but this effect is influenced by temperature. As the temperature approaches 80 F, germination decreases considerably irrespective of GA₃ treatment. Potato seed germination is also favored by low temperature, but constant low temperature alone does not insure complete germination of dormant seeds. More than 95% germination of dormant seeds can be obtained within a week either by application of GA₃ at low temperature or by diurnal alternation of temperature. A possible mechanism for control of seed germination is discussed.     

Variation in cardinal temperatures for germination among wheat (Triticum aestivum) genotypes

In most tropical regions where wheat is grown under irrigation, high temperatures at sowing adversely affect crop establishment and subsequent seedling survival. The objective of this study was to compare wheat (Triticum aestivum) genotypes for their ability to germinate and grow at high temperatures during the seedling stage. Twenty-five seeds each of 14 spring wheat cultivars were placed on moist filter paper at different temperatures (5°C to 40°C) in a one-way thermogradient plate to determine the cardinal temperatures for germination. Rate of germination at each temperature for each genotype was computed as the inverse of time taken for 50% of the seeds to germinate. Rate of germination for each genotype at different temperatures was modelled with temperature to determine the base (t_b), and optimum (t_opt) temperatures. Response of germination to temperature for each genotype was calculated as the slope of a linear regression of the rate of germination on temperature below t_opt. Genotypes differed in their optimum temperatures and Mexipak (= Kalyansona) had the lowest. Range in base temperature among the genotypes was between 0°C and 2°C differences but were not statistically significant though they might be biologically significant. Genotypes differed in their response to temperature with Gomam having the lowest rate, implying that it was slow to respond to increasing temperatures. Debeira and Cham 6 showed a similar response. Three lines which had performed well in spring wheat evaluation trials for moderate rainfall areas under heat stress had the highest response rate. It is concluded that combining higher optimum temperatures with faster response rates would result in better-adapted germplasm for regions where high temperatures persist at sowing.     

Dynamics of chromosomal instability in Welsh onion (Allium fistulosum L.): Influence of seed storage temperature

The age-related dynamics of chromosomal instability in cells of the root meristem of seedlings and germination capacity of seeds of Welsh onion (Allium fistulosum L.) in two storage temperature regimes in the course of six years following collection of the seeds are investigated. Seeds that had been stored at room temperature (14–28°C) lost germination capacity after six years of storage. The frequency of aberrant anaphases in these seeds grew from 2% in the first month of storage of the seeds to 80% in the 75th month. The germination capacity of seeds that had been stored at reduced temperatures (4–9°C) amounted to 73–77% in the sixth year, while the frequency of aberrant anaphases in these seeds remained within the range 2–4% throughout the six years. Thus, storage of Welsh onion seeds for six years at reduced temperatures tends to preserve the germination capacity of the seeds and prevents the development of chromosomal instability in the root meristem cells of the seedlings over this period.     

The germination of grass seeds after storage at different temperatures in aluminium foil and manilla paper packets

The germination of seeds of three species of forage grasses, Lolium perenne, Festuca pratensis and Dactylis glomerata, was studied after storage for 3–5 years under five different storage conditions: in aluminium foil packets at —25°C, 0°C and laboratory temperature (c. 18°C), and in manilla paper packets at 0°C and laboratory temperature. With Lolium perenne and Festuca pratensis high germination values at 3 and 7 days were obtained from seed stored at — 25 °C and 0°C in foil packets (5% moisture), but at laboratory temperatures, seed from foil packets gave lower germination values than those from manilla paper packets. At all three temperatures Dactylis glomerata germination after 7 and 14 days was higher in seed stored in foil than in manilla packages. With all three species stored in manilla packets, germination was higher after laboratory than cold storage.     

Low-temperature stratification strategies and growth regulators for rapid induction of Paris polyphylla var. yunnanensis seed germination

The seeds of Paris polyphylla var. yunnanensis are deeply dormant, and they remain dormant for 18 months or longer in their natural environment. Periodic exposure of the seeds to a low-temperature of 4 °C broke the dormancy in about 16 weeks (112 days). The most effective temperature stratification scheme was an interval of 14 days at 4 °C and 14 days at 22 °C. Both GA₃ and ethephon significantly enhanced the germination rate during the stratification treatment. The seed coat, particularly the mesophyll outer layer of the seed coat, strongly inhibited the germination. With removal of the seed coat and exposure of the uncoated seeds to 600 mg/l GA₃ for 48 h before the temperature stratification of 14 days at 4 °C and 14 days at 22 °C for 112 days, a germination percentage as high as 95.3% of the seeds was attained in about 160 days.     

Heat and smoke pre‐treatment of seeds to improve restoration of an endangered Mediterranean climate vegetation type

Invasive alien plants impact ecosystems, which often necessitates their removal. Where indigenous species recovery fails following removal alone, an active intervention involving reintroduction of seed of native species may be needed. This study investigated the potential for a combination of the fire cues of smoke and heat as a pre‐treatment of seeds in breaking dormancy and facilitating increased germination. Species were selected to represent different functional types within Cape Flats Sand Fynbos; a fire‐prone, critically endangered vegetation type in South Africa. Seeds were exposed to either a heat pulse (temperatures between 60 and 300°C for durations of between 30 s and 20 min) or dry after‐ripening (1 or 2 months at milder temperatures of 45°C or less). Thereafter, seeds were soaked in smoke solution for 18 h and subsequently placed on agar at 10/20°C for germination. Most species fell into one of two main groups: Seed germination in the first group was greatest following a lower temperature (60°C) heat pulse, an extended period of mild temperature (20/40°C or 45°C) exposure, or no pre‐treatment with heat. Seed germination in the second group was promoted after brief exposure to higher (100°C) temperatures. No germination occurred in any species following heat treatments of 150°C or higher. Species which responded better to higher temperatures were mainly those possessing physical dormancy, but seed morphology did not correlate with germination success. This study showed that heat stimulation of seeds is more widespread in fynbos plant families than previously known and will enable the development of better seed pre‐treatment protocols before large‐scale sowing as an active restoration treatment after alien plant clearing.     

Cryopreservation of conditionally dormant orthodox seeds of Betula pendula

To evaluate the feasibility of long-term cryopreservation of Polish provenances of silver birch (Betula pendula), the sensitivity of conditionally dormant seeds to extreme desiccation and/or the ultra-low temperature of liquid nitrogen (LN; ?196°C) was evaluated. The critical water content (WC) of desiccated seeds and the high-moisture freezing limit of seeds desiccated or moistened to various WCs and frozen for 24 h or for 2 years in LN was also determined. Germination tests revealed no critical WC for seeds [to 0.02 g H₂O g^?1 dry mass (g g^?1)]. Seeds tolerated freezing in LN within specific safe range of WC 0.02–0.23 g g^?1 (nuts). Seeds desiccated to the safe WC and stored in LN for 2 years had similar or higher germination as seeds stored at ?3°C for 2 years, depending on provenance. Therefore, long-term cryopreservation of B. pendula seeds in gene banks is feasible.